Abstract
The full-length cDNA corresponding to Stra8, a novel gene inducible by retinoic acid (RA) in P19 embryonal carcinoma cells, has been isolated and shown to encode a 45-kD protein. Both Stra8 mRNA and protein were induced in cells treated by all-trans and 9-cis retinoic acids. Two- dimensional gel analysis and dephosphorylation experiments revealed that the two stereoisomers of RA differentially regulate the phosphorylation status of the Stra8 protein, which was shown to exist in differently phosphorylated forms. Subcellular fractionation and immunocytochemistry studies showed that the Stra8 protein is cytoplasmic. During mouse embryogenesis, Stra8 expression was restricted to the male developing gonads, and in adult mice, the expression of Stra8 was restricted to the premeiotic germ cells. Thus, Stra8 protein may play a role in the premeiotic phase of spermatogenesis.
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- Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
- Bitgood M. J., Shen L., McMahon A. P. Sertoli cell signaling by Desert hedgehog regulates the male germline. Curr Biol. 1996 Mar 1;6(3):298–304. doi: 10.1016/s0960-9822(02)00480-3. [DOI] [PubMed] [Google Scholar]
- Bouillet P., Chazaud C., Oulad-Abdelghani M., Dollé P., Chambon P. Sequence and expression pattern of the Stra7 (Gbx-2) homeobox-containing gene induced by retinoic acid in P19 embryonal carcinoma cells. Dev Dyn. 1995 Dec;204(4):372–382. doi: 10.1002/aja.1002040404. [DOI] [PubMed] [Google Scholar]
- Bouillet P., Oulad-Abdelghani M., Vicaire S., Garnier J. M., Schuhbaur B., Dollé P., Chambon P. Efficient cloning of cDNAs of retinoic acid-responsive genes in P19 embryonal carcinoma cells and characterization of a novel mouse gene, Stra1 (mouse LERK-2/Eplg2). Dev Biol. 1995 Aug;170(2):420–433. doi: 10.1006/dbio.1995.1226. [DOI] [PubMed] [Google Scholar]
- Chambon P. A decade of molecular biology of retinoic acid receptors. FASEB J. 1996 Jul;10(9):940–954. [PubMed] [Google Scholar]
- Chambon P. The retinoid signaling pathway: molecular and genetic analyses. Semin Cell Biol. 1994 Apr;5(2):115–125. doi: 10.1006/scel.1994.1015. [DOI] [PubMed] [Google Scholar]
- Durand B., Saunders M., Leroy P., Leid M., Chambon P. All-trans and 9-cis retinoic acid induction of CRABPII transcription is mediated by RAR-RXR heterodimers bound to DR1 and DR2 repeated motifs. Cell. 1992 Oct 2;71(1):73–85. doi: 10.1016/0092-8674(92)90267-g. [DOI] [PubMed] [Google Scholar]
- Fyrberg E., Fyrberg C. C., Beall C., Saville D. L. Drosophila melanogaster troponin-T mutations engender three distinct syndromes of myofibrillar abnormalities. J Mol Biol. 1990 Dec 5;216(3):657–675. doi: 10.1016/0022-2836(90)90390-8. [DOI] [PubMed] [Google Scholar]
- Gaetano C., Matsumoto K., Thiele C. J. Retinoic acid negatively regulates p34cdc2 expression during human neuroblastoma differentiation. Cell Growth Differ. 1991 Oct;2(10):487–493. [PubMed] [Google Scholar]
- Gianni M., Zanotta S., Terao M., Garattini S., Garattini E. Effects of synthetic retinoids and retinoic acid isomers on the expression of alkaline phosphatase in F9 teratocarcinoma cells. Biochem Biophys Res Commun. 1993 Oct 15;196(1):252–259. doi: 10.1006/bbrc.1993.2242. [DOI] [PubMed] [Google Scholar]
- Green S., Issemann I., Sheer E. A versatile in vivo and in vitro eukaryotic expression vector for protein engineering. Nucleic Acids Res. 1988 Jan 11;16(1):369–369. doi: 10.1093/nar/16.1.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gronowski A. M., Bertics P. J. Modulation of epidermal growth factor receptor interaction with the detergent-insoluble cytoskeleton and its effects on receptor tyrosine kinase activity. Endocrinology. 1995 May;136(5):2198–2205. doi: 10.1210/endo.136.5.7720669. [DOI] [PubMed] [Google Scholar]
- HOWELL J. M., THOMPSON J. N., PITT G. A. Histology of the lesions produced in the reproductive tract of animals fed a diet deficient in vitamin A alcohol but containing vitamin A acid. I. The male rat. J Reprod Fertil. 1963 Apr;5:159–167. doi: 10.1530/jrf.0.0050159. [DOI] [PubMed] [Google Scholar]
- Hacker A., Capel B., Goodfellow P., Lovell-Badge R. Expression of Sry, the mouse sex determining gene. Development. 1995 Jun;121(6):1603–1614. doi: 10.1242/dev.121.6.1603. [DOI] [PubMed] [Google Scholar]
- Heath J. K., Suva L. J., Yoon K., Kiledjian M., Martin T. J., Rodan G. A. Retinoic acid stimulates transcriptional activity from the alkaline phosphatase promoter in the immortalized rat calvarial cell line, RCT-1. Mol Endocrinol. 1992 Apr;6(4):636–646. doi: 10.1210/mend.6.4.1584226. [DOI] [PubMed] [Google Scholar]
- Heyman R. A., Mangelsdorf D. J., Dyck J. A., Stein R. B., Eichele G., Evans R. M., Thaller C. 9-cis retinoic acid is a high affinity ligand for the retinoid X receptor. Cell. 1992 Jan 24;68(2):397–406. doi: 10.1016/0092-8674(92)90479-v. [DOI] [PubMed] [Google Scholar]
- Kastner P., Mark M., Chambon P. Nonsteroid nuclear receptors: what are genetic studies telling us about their role in real life? Cell. 1995 Dec 15;83(6):859–869. doi: 10.1016/0092-8674(95)90202-3. [DOI] [PubMed] [Google Scholar]
- Kastner P., Mark M., Leid M., Gansmuller A., Chin W., Grondona J. M., Décimo D., Krezel W., Dierich A., Chambon P. Abnormal spermatogenesis in RXR beta mutant mice. Genes Dev. 1996 Jan 1;10(1):80–92. doi: 10.1101/gad.10.1.80. [DOI] [PubMed] [Google Scholar]
- Kato M., Sung W. K., Kato K., Goodman D. S. Immunohistochemical studies on the localization of cellular retinol-binding protein in rat testis and epididymis. Biol Reprod. 1985 Feb;32(1):173–189. doi: 10.1095/biolreprod32.1.173. [DOI] [PubMed] [Google Scholar]
- Kemp B. E., Pearson R. B. Protein kinase recognition sequence motifs. Trends Biochem Sci. 1990 Sep;15(9):342–346. doi: 10.1016/0968-0004(90)90073-k. [DOI] [PubMed] [Google Scholar]
- Koopman P., Münsterberg A., Capel B., Vivian N., Lovell-Badge R. Expression of a candidate sex-determining gene during mouse testis differentiation. Nature. 1990 Nov 29;348(6300):450–452. doi: 10.1038/348450a0. [DOI] [PubMed] [Google Scholar]
- Koshimizu U., Watanabe M., Nakatsuji N. Retinoic acid is a potent growth activator of mouse primordial germ cells in vitro. Dev Biol. 1995 Apr;168(2):683–685. doi: 10.1006/dbio.1995.1113. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Leid M., Kastner P., Chambon P. Multiplicity generates diversity in the retinoic acid signalling pathways. Trends Biochem Sci. 1992 Oct;17(10):427–433. doi: 10.1016/0968-0004(92)90014-z. [DOI] [PubMed] [Google Scholar]
- Levin A. A., Sturzenbecker L. J., Kazmer S., Bosakowski T., Huselton C., Allenby G., Speck J., Kratzeisen C., Rosenberger M., Lovey A. 9-cis retinoic acid stereoisomer binds and activates the nuclear receptor RXR alpha. Nature. 1992 Jan 23;355(6358):359–361. doi: 10.1038/355359a0. [DOI] [PubMed] [Google Scholar]
- Levy E., Liem R. K., D'Eustachio P., Cowan N. J. Structure and evolutionary origin of the gene encoding mouse NF-M, the middle-molecular-mass neurofilament protein. Eur J Biochem. 1987 Jul 1;166(1):71–77. doi: 10.1111/j.1432-1033.1987.tb13485.x. [DOI] [PubMed] [Google Scholar]
- Lufkin T., Dierich A., LeMeur M., Mark M., Chambon P. Disruption of the Hox-1.6 homeobox gene results in defects in a region corresponding to its rostral domain of expression. Cell. 1991 Sep 20;66(6):1105–1119. doi: 10.1016/0092-8674(91)90034-v. [DOI] [PubMed] [Google Scholar]
- Lufkin T., Lohnes D., Mark M., Dierich A., Gorry P., Gaub M. P., LeMeur M., Chambon P. High postnatal lethality and testis degeneration in retinoic acid receptor alpha mutant mice. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7225–7229. doi: 10.1073/pnas.90.15.7225. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mangelsdorf D. J., Thummel C., Beato M., Herrlich P., Schütz G., Umesono K., Blumberg B., Kastner P., Mark M., Chambon P. The nuclear receptor superfamily: the second decade. Cell. 1995 Dec 15;83(6):835–839. doi: 10.1016/0092-8674(95)90199-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Manova K., Nocka K., Besmer P., Bachvarova R. F. Gonadal expression of c-kit encoded at the W locus of the mouse. Development. 1990 Dec;110(4):1057–1069. doi: 10.1242/dev.110.4.1057. [DOI] [PubMed] [Google Scholar]
- Oulad-Abdelghani M., Bouillet P., Chazaud C., Dollé P., Chambon P. AP-2.2: a novel AP-2-related transcription factor induced by retinoic acid during differentiation of P19 embryonal carcinoma cells. Exp Cell Res. 1996 Jun 15;225(2):338–347. doi: 10.1006/excr.1996.0184. [DOI] [PubMed] [Google Scholar]
- Rajan N., Sung W. K., Goodman D. S. Localization of cellular retinol-binding protein mRNA in rat testis and epididymis and its stage-dependent expression during the cycle of the seminiferous epithelium. Biol Reprod. 1990 Nov;43(5):835–842. doi: 10.1095/biolreprod43.5.835. [DOI] [PubMed] [Google Scholar]
- Rochette-Egly C., Gaub M. P., Lutz Y., Ali S., Scheuer I., Chambon P. Retinoic acid receptor-beta: immunodetection and phosphorylation on tyrosine residues. Mol Endocrinol. 1992 Dec;6(12):2197–2209. doi: 10.1210/mend.6.12.1283441. [DOI] [PubMed] [Google Scholar]
- Rochette-Egly C., Lutz Y., Saunders M., Scheuer I., Gaub M. P., Chambon P. Retinoic acid receptor gamma: specific immunodetection and phosphorylation. J Cell Biol. 1991 Oct;115(2):535–545. doi: 10.1083/jcb.115.2.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roy B., Taneja R., Chambon P. Synergistic activation of retinoic acid (RA)-responsive genes and induction of embryonal carcinoma cell differentiation by an RA receptor alpha (RAR alpha)-, RAR beta-, or RAR gamma-selective ligand in combination with a retinoid X receptor-specific ligand. Mol Cell Biol. 1995 Dec;15(12):6481–6487. doi: 10.1128/mcb.15.12.6481. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rudnicki M. A., Ruben M., McBurney M. W. Regulated expression of a transfected human cardiac actin gene during differentiation of multipotential murine embryonal carcinoma cells. Mol Cell Biol. 1988 Jan;8(1):406–417. doi: 10.1128/mcb.8.1.406. [DOI] [PMC free article] [PubMed] [Google Scholar]
- THOMPSON J. N., HOWELL J. M., PITT G. A. VITAMIN A AND REPRODUCTION IN RATS. Proc R Soc Lond B Biol Sci. 1964 Feb 18;159:510–535. doi: 10.1098/rspb.1964.0017. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wang S. Y., LaRosa G. J., Gudas L. J. Molecular cloning of gene sequences transcriptionally regulated by retinoic acid and dibutyryl cyclic AMP in cultured mouse teratocarcinoma cells. Dev Biol. 1985 Jan;107(1):75–86. doi: 10.1016/0012-1606(85)90377-x. [DOI] [PubMed] [Google Scholar]